Cell Signaling Technology
α-Tubulin Antibody
2144
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Rab proteins constitute the largest family of monomeric small GTPases that oscillate between active (GTP- bound) and inactive (GDP- bound) conformations. The spatio-temporal activation of Rabs is controlled by the GDP-GTP exchange factors (GEFs), mediating their activation, and the GTP hydrolysis activating proteins (GAPs), terminating their activity. Rab GTPases execute diverse tasks in membrane trafficking by driving cargo collection into nascent transport vesicles, linking vesicles to cytoskeletal elements, recruiting tethers, and finally docking vesicles to the target compartment, leading to membrane fusion. The goal of this project is to characterize a specific GEF for Golgi localized Rab6 GTPase, and to analyze its effect in Rab6-mediated processes at the Golgi complex. In the present experiment, alpha-tubulin antibody was used as a loading control for the gel. This antibody was recommended by a researcher from a neighboring lab.
Western Blot
Whole cell extracts of HeLa cells
1:5000 dilution in blocking buffer; 1 hr at room temp
5% Milk powder in 1xTBS-T(0.05%)
Goat-anti Rabbit-IgG-HRP conjugated; 1:10000 dilution in blocking buffer; 1 hr at room temp
None
ECL regular; 2 sec exposure on a film
The loading of samples was uniform across three wells as judged by alpha-tubulin intensity.
Specific signal
Alpha-tubulin antibody from Cell Signaling is a perfect antibody for Western Blotting.